Exploring the feasibility of Whole Genome Amplification (WGA) for recovering ancient DNA

The Leonardo da Vinci DNA project aims to recover and analyse the DNA of Leonardo da Vinci (LdV). One of the approaches to this end is to obtain his DNA through metagenomic studies from his works of art, especially drawings and codices. Metagenomic studies were already performed on LdV's drawings using the Nanopore sequencing technology, but these studies focused on the microbiomes, in order to preserve these artworks from biodeterioration and not on the human DNA also detected.

In this feasibility study, one of the tasks foreseen is to provide the "Leonardo da Vinci DNA Project" with the part of the data related to the human DNA sequences obtained in the above-mentioned study for further analysis. However, the major objective in this study is to test a protocol using the combined strategy of DNA amplification with Phi 29 polymerase and sequencing with Nanopore platform and to explore whether this strategy could be applied/ adapted to amplify and sequence ancient (aDNA) when present in a mixture of environmental (eDNA).

To address this challenge, we will focus on a specific material: the parchment, as its written information represents a historical, artistic and social documentary source. In addition, the physical object of the parchment contains a considerable amount of biological information (bioarchive) containing environmental (eDNA) and ancient DNA (aDNA), which makes this material the perfect candidate for the analysis of both at the same time. The amount of DNA obtained in biocodicological studies is minimal, due to the obligatory non-invasive sampling on parchment, which makes subsequent steps for analysis difficult. Therefore, the application of the Phi 29 strategy would also represent a step forward in the field of biocodicology and in the field of ancient DNA in general.